Promoting the accumulation of tumor-specific T cells in tumor tissues by dendritic cell vaccines and chemokine-modulating agents

• Published in: Nature protocols Vol. 13; no. 2; pp. 335 - 357
• Main Authors: Obermajer, Nataša, Urban, Julie, Wieckowski, Eva, Muthuswamy, Ravikumar, Ravindranathan, Roshni, Bartlett, David L, Kalinski, Pawel
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01-02-2018; Nature Publishing Group
• Subjects: 13/31; 631/1647/664; 631/250/1619/554/1834/1269; 631/250/251; 631/67/580; 692/4028/67/1059/4042; Accumulation; Adjuvants; Analytical Chemistry; Animal tissues; Animals; Biological Techniques; Cancer treatment; Cancer Vaccines - chemical synthesis; Cancer Vaccines - metabolism; Cell Line; Chemokines; Combinatorial analysis; Combined vaccines; Computational Biology/Bioinformatics; Cytotoxicity; Dendritic cells; Dendritic Cells - physiology; Dendritic structure; Enzyme-linked immunosorbent assay; Health aspects; Homing; Homing behavior; Immunity; Immunogenicity; Immunotherapy; Immunotherapy - methods; Implantation; Innovations; Life Sciences; Lymph nodes; Lymph Nodes - immunology; Lymphocytes; Lymphocytes T; Mice; Mice, Inbred C57BL; Microarrays; Microenvironments; Neoplasms; Organic Chemistry; Organs; Production methods; protocol; Spleen - immunology; T cell receptors; T-Lymphocytes, Cytotoxic - immunology; T-Lymphocytes, Cytotoxic - physiology; Tumor microenvironment; Tumor Microenvironment - physiology; Tumors; Vaccines
• ISSN: 1754-2189; 1750-2799
• DOI: 10.1038/nprot.2017.130

This protocol describes how to generate and use dendritic cells and combinatorial adjuvants to modulate tumor microenvironment and increase T-cell accumulation in tumors
in vivo
.
This protocol describes how to induce large numbers of tumor-specific cytotoxic T cells (CTLs) in the spleens and lymph nodes of mice receiving dendritic cell (DC) vaccines and how to modulate tumor microenvironments (TMEs) to ensure effective homing of the vaccination-induced CTLs to tumor tissues. We also describe how to evaluate the numbers of tumor-specific CTLs within tumors. The protocol contains detailed information describing how to generate a specialized DC vaccine with augmented ability to induce tumor-specific CTLs. We also describe methods to modulate the production of chemokines in the TME and show how to quantify tumor-specific CTLs in the lymphoid organs and tumor tissues of mice receiving different treatments. The combined experimental procedure, including tumor implantation, DC vaccine generation, chemokine-modulating (CKM) approaches, and the analyses of tumor-specific systemic and intratumoral immunity is performed over 30–40 d. The presented ELISpot-based
ex vivo
CTL assay takes 6 h to set up and 5 h to develop. In contrast to other methods of evaluating tumor-specific immunity in tumor tissues, our approach allows detection of intratumoral T-cell responses to nonmanipulated weakly immunogenic cancers. This detection method can be performed using basic laboratory skills, and facilitates the development and preclinical evaluation of new immunotherapies.